Differential absorption flattening optical effects are significant in the circular dichroism spectra of large membrane fragments

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Abstract
The circular dichroism spectra of membrane particles are distorted by effects of differential absorption flattening, which are a consequence of the nonrandom distribution of chromophores in these samples. We have shown that this phenomenon is not significant in small unilamellar vesicles with high lipid to protein ratios [Mao, D., and Wallace, B. A. (1984) Biochemistry 23, 2667-2673]. It has recently been claimed [Glaeser, R. M., and B. K. (1985) Biochemistry 24, 6398-6401] that absorption flattening effects are also inconsequential in large membrane fragments with high protein concentrations, such as purple membrane sheets. This paper will demonstrate that absorption flattening is significant in these samples and that it causes substantial distortion of the calculated protein secondary structures derived from the uncorrected circular dichroism data.